The invention is concerned with a melon constitutive triple response (CTR1) homologue, called mCTR, vectors, cells and transgenic plants which comprise the coding sequence for mCTR or a biologically active fragment thereof and methods of producing transgenic plants which express mCTR or a biologically active fragment thereof.
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Ethylene (CH2xe2x95x90CH2), is a naturally occurring plant hormone which has diverse effects on plant growth and development when produced by the plant itself or applied exogenously. Ethylene promotes senescence in plants, both in selected groups of cells and in whole organs such as fruits, leaves and flowers. Ethylene mediated effects, such as the stimulation of ripening in fruits and vegetables; leaf abscission, yellowing and epinasty, and fading and wilting in flowers are of considerable commercial importance to fruit, vegetable and flower-related industries. Large quantities of ethylene are produced by plants during ripening and senescence and in response to trauma caused by chemicals, temperature extremes, water stress, ultraviolet light, insect damage, disease and mechanical wounding.
Reducing ethylene biosynthesis is an effective way of prolonging the ripening process in fruit and also has potential to delay post harvest senescence. An alternative to reducing ethylene biosynthesis is to reduce the plant""s ability to perceive and respond to ethylene by interfering with the signal transduction pathway leading to the ethylene response. A plant""s ability to respond to ethylene is correlated with transcription of ethylene perception pathway components (Payton et al., 1996), and mutant plants insensitive to ethylene show a delayed senescence phenotype (Oh et al. 1997; Chao et al. 1997). Great strides have been made in elucidating the signal transduction cascade responsible for ethylene perception in plants.
The ethylene receptor and downstream components of the ethylene perception pathway have been identified, and in some plant species the corresponding genes have been cloned. In Arabidopsis, the ethylene receptor is a member of an extended gene family which includes ETR1, ERS1, ERS2, ETR2, and EIN4 (Hua et al., 1998). Mutations at these loci result in plants that are insensitive to ethylene, and the mutations studied to date have been dominant.
A need exists for further elucidating the effects of blocking the ethylene response on fruit, vegetable and flower development and for modulating the ethylene response in a variety of agricultural industries.
It is a general object of the invention to provide methods for interference with ethylene perception and ethylene response in the fruit of fruit-bearing plants.
The invention provides an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a melon protein with the activity of a constitutive triple response (CTR) protein, wherein the nucleic acid sequence is selected from the group consisting of:
(a) a nucleic acid sequence encoding a protein comprising the amino acid sequence of SEQ ID NO: 2;
(b) a nucleic acid sequence that is SEQ ID NO: 1;
(c) a nucleic acid sequence that is nucleotides A-3286 of SEQ ID NO:1, wherein A is any one of nucleotides 1440-1444;
(d) a nucleic acid sequence that has at least 80, 85 or 90% sequence identity to the coding region of (a), (b) or (c)
(e) a nucleic acid sequence that will hybridize under moderate to high stringency conditions to the sequence presented as SEQ ID NO:1, or the complement thereof;
(f) a fragment of the nucleic acid sequence of (a), (b) or (c) wherein the fragment encodes a protein which has the activity of a constitutive triple response (CTR) protein; and
(g) a nucleic acid sequence that is degenerate as a result of the genetic code to the nucleic acid sequence of (a), (b), (c), (d), (e) or (f).
In a related aspect, the invention provides a plant expression vector comprising an mCTR nucleic acid sequence selected from those described above, operably linked to control sequences recognized by a plant cell transformed with the vector and transgenic plant cells comprising the nucleic acid sequence or plant expression vector.
The invention also provides mature transgenic plants comprising such transgenic plant cells.
In a further related aspect, the invention provides an isolated protein having the activity of a constitutive triple response (CTR) protein, wherein the protein is encoded by a nucleic acid sequence selected from those described above.
A method for producing transgenic plant lines having a decreased response to ethylene is also provided by the invention. The method includes the steps of introducing a plant expression vector, comprising an mCTR nucleic acid sequence selected from those described above, into cells of a plant under conditions effective to yield transformed plant cells, selecting for transformed plant cells and growing the selected plant cells to produce a transgenic plant line, the seedlings of which exhibit a modulated triple response to ethylene.
These and other objects and features of the invention will be more fully appreciated when the following detailed description of the invention is read in conjunction with the accompanying drawings.